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Biomedical subjects

J R Stephenson

Publications and source records attributed to J R Stephenson.

At least 181 records · Page 10Linked to original sources

Feline oncornavirus-associated cell membrane antigen: evidence for an immunologically crossreactive feline sarcoma virus-coded protein.

The feline oncornavirus-associated cell membrane antigen (FOCMA) acts as a target for natural immuno-surveillance against tumor development in the cat. In the present study, mink and rat cells nonproductively transformed by feline sarcoma virus (FeSV) were shown to express FOCMA as well as 5'-terminal feline leukemia virus (FeLV) gag gene proteins, p15 and p12. In contrast, such cells lack detectable levels of other FeLV gag gene-coded proteins or the env gene product, gp70. FOCMA, p15, and p12 antigen expression is initially in the form of an 80,000-100,000 molecular weight precursor which, upon post-translational cleavage, gives rise to a 65,000 molecular weight component that contains FOCMA and a 25,000 molecular weight component containing p15 and p12. Feline lymphoma cells, including those from several tumors that lacked detectable levels of FeLV structural protein expression, were shown to be FOCMA-positive. These findings strongly suggest that FOCMA represents an FeSV-coded transformation specific protein and provide preliminary information regarding the position within the FeSV genome coding for its synthesis.

Animals↗

Feline oncornavirus-associated cell-membrane antigen (FOCMA): distinction between FOCMA and the major virion glycoprotein.

The humoral antibody response of feline leukemia virus (FeLV)-exposed cats to the feline oncornavirus-associated tumor cell-membrane antigen (FOCMA) is directly correlated with immunosurveillance against tumor development under natural conditions. By means of membrane immunofluorescence and radioimmunoprecipitation, the antibody response to FOCMA was found to be independent of the antibody response to the major envelope and core proteins of FeLV, gp70 and p30. This was especially true for healthy viremic cats, where antigenemia with circulating FeLV gp70 and p30 apparently binds any free antibody to these proteins, but high levels of FOCMA antibody are often concurrently present. Exhaustive in vitro absorption of highly immune nonviremic serum with gp70 and p30 also failed to remove FOCMA antibody activity. These results indicate that FOCMA is not one of these major FeLV structural proteins.

Animals↗

Immunoprevention of x-ray-induced leukemias in the C57BL mouse.

An attempt to prevent irradiation-induced thymic lymphomas in C57BL mice was made by inducing active immunity to endogenous type-C virus with inactivated Rauscher murine leukemia virus (MuLV) or inactivated Gross MuLV or by transferring passive immunity to endogenous type-C virus with goat anti-Gross MuLV IgG. Control groups received the following immunogen or treatment: inactivated simian sarcoma virus, complete Freund's adjuvant, normal goat IgG, and diluent, in both irradiated and nonirradiated C57BL mice Active immunity to the 70,000 molecular weight glycoprotein AKR-gp70 by immunization with Rauscher MuLV and passive immunity to AKR-gp70 by passive transfer of goat anti-Gross-MuLV IgG was measurable throughout some of the latent period of tumor development; in these two groups a significant reduction in tumor incidence was observed, as compared to the other experimental and control groups. Thus, the present findings support the concept of a type-C virus etiology of irradiation-induced leukemias and demonstrate the applicability of immunologic techniques directed against the endogenous type-C virus in the prevention of this disease.

Animals↗

Segregation of genetic information for a B-tropic leukemia virus with the structural locus for BALB:virus-1.

A B-tropic type-C RNA virus isolatable from lymphoreticular tumors of the inbred BALB/c mouse strain has previously been shown to be leukemogenic in its natural host. This virus is not chemically inducible from BALB/c embryo cells or from embryo lines containing segregating inducibility loci for two known endogenous type-C viruses of BALB/c cells. Molecular hybridization and type-specific immunologic assays demonstrate a high degree of genetic homology between the B-tropic leukemia virus and BALB:virus-1, an N-tropic endogenous virus of BALB/c cells. Genetic sequences specific for BALB:virus-1 are shown to segregate with the locus for BALB:virus-1 induction in genetic crosses between BALB/c and the noninducible NIH Swiss strain. Thus, if the information of the B-tropic virus is encoded in the genome of the animal, it must be closely linked to the structural locus for BALB:virus-1. The evidence is consistent with a mechanism by which a small genetic alteration in BALB:virus-1 leads to a virus, whose growth is unrestricted, and subsequently to the development of neoplasia.

Animals↗

Characterization of virus-specific messenger RNAs from avian fibroblasts infected with fowl plague virus.

In cell-free protein synthesizing systems from wheat embryos, messenger RNAs extracted from chick embryo fibroblasts infected with fowl plague virus direct the synthesis of nine virus-specific polypeptides, two of which may be related to the virus-specific glycopolypeptides. All of the mRNAs are complementary in sequence to virion RNA, and RNAs which do not contain poly A appear to be translated as efficiently as their polyadenylated counterparts. Under certain conditions of incubation, virion RNA also directs the synthesis of discrete polypeptides but these products are not detected in infected cells.

Animals↗

Distribution and expression in mammals of genes ralated to an endogenous type C RNA virus of Odocoileus hemionus.

An endogenous type C virus recently isolated from the Columbian black-tailed deer (Odocoileus hemionus) was used as a molecular probe to study the distribution of virus-related nucleotide sequences in cellular DNAs of mammalian species. By DNA-DNA hybridization, the most extensive homology was demonstrated between the viral complementary DNA and cellular DNA isolated from Odocoileus species. DNAs of representatives of other genera within the same family, Cervidae, were partially related to the virus, consistent with the phylogenetic relationship of these species to Odocoileus. O. hemionus viral sequences were also detected within cellular DNAs of members of a more distantly related artiodactyl family, Bovidae. These findings suggest the genetic transmission of type C viral genes within cervids and bovids for a period of at least 25 to 30 million years. There was no detectable nucleotide sequence homology between O. hemionus virus and representatives of other major groups of mammalian type C viruses. These results indicate that despite the known antigenic relatedness of mammalian type C viruses, the O. hemionus virus has diverged sufficiently to be considered the prototype of a separate group. By radioimmunological techniques, it was possible to detect and partially purify, from normal tissues of cervid species, antigens related to the major structural protein of the O. hemionus virus. The present findings, that O. hemionus virus has been genetically transmitted for millions of years and yet has maintained the ability to be expressed as infectious virus, argue for positive evolutionary selective pressures for the maintenance of type C viral genes.

Animals↗

Type C viral gag gene expression in chicken embryo fibroblasts and avian sarcoma virus-transformed mammalian cells.

Sensitive radioimmunoassays were developed for avian type C viral gag gene-coded proteins. These assays were used to examine the restriction to virus production by avian embryo cells and mammalian cells transformed by avian sarcoma viruses. The results indicate that although a high-molecular-weight primary translational product of the gag gene is expressed, its cleavage and processing are incomplete. Furthermore, analysis of intermediate cleavage products provided information regarding the order of sequences coding for the individual viral proteins within the avian type C viral gag gene.

Alpharetrovirus↗

Biochemical and immunological characterization of the major envelope glycoprotein of bovine leukemia virus.

The major envelope glycoprotein of bovine leukemia virus was isolated by lectin-bound Sepharose and DEAE-cellulose column chromatography. This protein was shown to have a molecular weight of about 41,000 and to lack detectable immunological cross-reactivity with glycoproteins of other oncornaviruses. Sera obtained from 100% of cattle examined with clinically diagnosed lymphosarcoma contained high-titered antibody to 125I-labeled bovine leukemia virus glycoprotein, whereas sera from animals in a disease-free herd were antibody negative.

Animals↗

Endogenous RD-114 virus genome expression in malignant tissues of domestic cats.

Endogenous xenotropic cat type C virus (RD-114)- and infectious feline leukemia virus (FeLV)-specific gene expressions were measured in spontaneous sarcomas carcinomas, and nonmalignant cat tissues by molecular hybridization for virus-specific RNA and competition radio-immunoassays for the major internal protein (p30) of these two viruses. The results indicate that RD-114 gene expression in sarcomas and carcinomas at both RNA and p30 levels is significantly higher than histologically normal tissues from cats free of cancer. In contrast, the levels of FeLV viral RNA and p30 are fount to be low or undetectable in the majority of these tumored and normal tissues examined. Whereas variability in the amounts of RD-114 OR FeLV RNA and p30 expressed is found in tissues from different cats, their expression is fairly uniform in multiple malignant tissues of the same cat. The finding of widespread occurrence of elevated RD-114 gene expression in sarcomas and carcinomas is consistent with our similar observation with natural lymphomas of domestic cats and suggests that expression of certain functions of this endogenous virus may be etiologically involved in the development of many different spontaneous neoplasms of cats.

Animals↗

Feline leukemia virus: biochemical and immunological characterization of gag gene-coded structural proteins.

The major non-glycosylated structural proteins of feline leukemia virus have been isolated, and competition immunoassays have been developed for each. These proteins include the 27,000- to 30,000-molecular-weight major internal antigen designated p30, a 15,000-molecular-weight protein (p15), an acidic protein of 12,000 molecular weight (p12), and a highly basic 10,000-molecular-weight protein (p10). Immunologically and biochemically corresponding proteins of feline and murine leukemia viruses have been identified. and, on the basis of analogy to the known sequence of a prototype type C virus of mouse origin, the map order of the gag region of the feline type C viral genome has been tentatively deduced as NH2-p15-p12-p10-COOH. The demonstration of two feline leukemia virus gag gene-coded proteins, p15 and p12, expressed in the form of an uncleaved precursor in a mink cell line nonproductively transformed by feline sarcoma virus provides indirect support for the proposed sequence.

AKR murine leukemia virus↗

Genetic factors infuencing mouse type-C RNA virus induction by naturally occurring B cell mitogens.

The present studies demonstrate that xenotropic type C virus is efficiently released in response to lipopolysaccharide by spleen cells of a wide variety of inbred mouse strains. Lipopolysaccharide-mediated virus release primarily involves B lymphocytes and is in part genetically determined. Virus release can also be efficiently stimulated by other naturally occurring B cell mitogens, including Nocardia water soluble mitogen, and PPD. The evidence indicates that these agents act synergistically with halogeneated pyrimidines, but not with each other, to cause virus release. These results indicate that B cell mitogens act to release virus by a mechanism that differs from that of halogenated pyrimidines.

Animals↗

Evaluation of radioimmunoprecipitation for the detection of bovine leukemia virus infection in domestic cattle.

The potential application of a recently developed radioimmunoprecipitation test for antibody directed against the major structural protein of bovine leukemia virus was evaluated for use in detection of BLV-infection in domestic cattle. This technique was found to be considerably more sensitive than serologic procedures currently being utilized for this purpose. Radioimmunoprecipitation was also shown to have distinct advantages as compared to hematologic criteria, such as specified by Bendixen's index, for identification of BLV-infected animals. By the use of radioimmunoprecipitation, high levels of antibody to BLV were demonstrated in sera of animals with confirmed adult lymphosarcoma, but not in animals with a less common sporadic form of the disease which occurs in calves.

Aging↗

Bovine lymphosarcoma: development of a radioimmunologic technique for detection of the etiologic agent.

A highly sensitive and specific radioimmunoassay has been developed for the major structural protein of an oncornavirus etiologically associated with bovine lymphosarcoma. This test can be used to identify cattle which have been exposed to the bovine leukemia virus and may thus develop or transmit the disease. Analysis of randomly obtained serums indicates that infection that infection with this virus is widespread among cattle.

Animals↗

Structural polypeptides of mammalian type C RNA viruses. Isolation and immunologic characterization of a low molecular weight polypeptide, p10.

Low molecular weight polypeptides of several mammalian type C RNA tumor viruses were purified by sequential ion exchange chromatography and molecular sizing techniques. These included a polypeptide with a molecular weight of 10,000 to 11,000, p 10, from two type C viruses of mouse origin. Rauscher- and Moloney-murine leukemia virus (MuL virus), and from an infectious type C virus isolate of the woolly monkey. The p12 structural polypeptides of these viruses as well as Rauscher-MuL virus p15 were also purified. By using radioimmunoassays developed for each polypeptide, it was possible to demonstrate that all three low molecular weight polypeptides, p15, p12, and p10, were immunologically unique. Among type C viral structural polypeptides, p10 has been least well characterized immunologically. The results of the present study indicate that p10 is virus-coded and possesses strong group-specific antigenic determinants. By use of appropriate immunoassays, broadly reactive interspecies determinants shared by mammalian type C virus isolates of murine, feline, and primate origin, were also demonstrated. The interspecies antigenic determinants of p10 were shown to be as broadly cross-reactive as those exhibited by the major type C virus structural polypeptide, p30.

Antigen-Antibody Reactions↗

gag Gene of mammalian type-C RNA tumour viruses.

The translation product of the gag gene of mammalian type-c- RNA viruses is a 65,000-68,000 molecular weight precursor polypeptide (Pr65) whose cleavage leads to the formation of four virion proteins, p30, p15, p12 and p10. An immunological approach has been used to establish the arrangement of the sequences coding for these proteins within the viral genome as (5') p15-p12-p30-p10 (3').

Chromosome Mapping↗